Figure 7.

JAZ8 Represses the Interaction between RHD6 and RSL1.

(A) Yeast two-hybrid assays showing the interaction between RHD6 and RSL1, and that the N-terminal part of RHD6 (RHD6^1-193) is responsible for the interaction. Yeast cells were plated onto dropout medium lacking Leu, Trp, His, and Ade and containing 3 mM 3-aminotriazole and observed after 2 d. BD, DNA binding domain; AD, Activation domain.

(B) BiFC analyses showing that RHD6 interacts with RSL1 in plant cells. Fluorescence was observed in nuclei of transformed tobacco cells, resulting from the complementation of RHD6-nYFP with RSL1-cYFP. No signal was obtained from negative controls in which RHD6-nYFP was coexpressed with RSL1^200-307-cYFP (the C-terminal domain of RSL1 fused with cYFP), or RHD6^194–298-nYFP (the C-terminal domain of RHD6 fused with nYFP) was coexpressed with RSL1-cYFP. Nuclei are indicated by DAPI staining. Bar = 15 μm.

(C) BiFC analyses showing that JAZ8 interferes with the interaction between RHD6 and RSL1. Similar results were obtained from at least three independent replicates. Bar = 30 μm.

(D) Quantitative analysis of YFP fluorescence intensity in (C). Fifty independent fluorescent spots were valued for fluorescence intensity. Error bars represent SD. Bars with different letters are significantly different from each other (P < 0.05). Data were analyzed by ANOVA.